Heart transplantation encompasses a wide range of issues, including basic medical and immunological research, ethics, and sophisticated clinical strategies, with prolonged heart preservation and prevention of ischemia/reperfusion injury being the most important issues. The heart preservation strategy is closely related to myocardial protection in routine cardiac surgery, which has a long history of development. In this article, our efforts at Osaka University were reviewed from the aspects of developing myocardial protection and strategy for vulnerable targets such as neonatal hearts, preoperatively damaged hearts, left ventricular hypertrophy, and in heart transplantation. Recognizing the role of activated neutrophils in reperfusion injury, we developed the system of leukocyte-depleted terminal cardioplegia. This strategy has also applied to clinical heart transplantation. On the otherhand, during the period before the resumption of heart transplantation from brain-dead donors, we conducted experimental studies for the possible clinical start of heart transplantation from donors after cardiac death (non-heart-beating donor) covering 24-hour cold storage. In summary, we hope that our efforts described here can contribute to the current practice of myocardial preservation in heart transplantation.